Anatomy and cell wall ultrastructure of sunflower stalk rind

The anatomy and ultrastructure of sunflower stalk rind are closely related to its conversion and utilization. We studied systematically the anatomy and ultrastructure of the stalk rind using light, scanning electron, transmission electron and fluorescence microscopy. The results showed that the stalk rind consisted of phloem fibers (PF), xylem fibers (XF), vessel elements (V), ground parenchyma cells (GPC), axial parenchyma cells (APC), xylem ray parenchyma cells (XRPC), and pith ray parenchyma cells (PRPC). These cell walls were divided into the middle lamella, primary wall, and secondary wall (S). It was found that the S of PF, XF and V was further divided into three layers (S1–S3), while the S of APC, GPC, XRPC and PRPC showed a non-layered cell wall organization or differentiated two (S1, S2) to seven layers (S1–S7). Our research revealed the plasmodesmata characteristics in the pit membranes (PMs) between parenchyma cells (inter-GPCs, inter-XRPCs, and inter-PRPCs). The morphology of the plasmodesmata varied with the types of parenchyma cells. The thickness and diameter of PMs between the cells (inter-Vs, V–XF, V–APC, and V–XRPC) were greater than that of PMs between parenchyma cells. The cell corners among parenchyma cells were intercellular space. The lignification degree of vessels was higher than that of parenchyma cells and fibers. The results will provide useful insights into the biological structure, conversion and utilization of sunflower stalk rind.

Cell Wall Layer Induced in Xylem Fibers of Flax Upon Gravistimulation Is Similar to Constitutively Formed Cell Walls of Bast Fibers

In the fibers of many plant species after the formation of secondary cell walls, cellulose-enriched cell wall layers (often named G-layers or tertiary cell walls) are deposited which are important in many physiological situations. Flax (Linum usitatissimum L.) phloem fibers constitutively develop tertiary cell walls during normal plant growth. During the gravitropic response after plant inclination, the deposition of a cellulose-enriched cell wall layer is induced in xylem fibers on one side of the stem, providing a system similar to that of tension wood in angiosperm trees. Atomic force microscopy (AFM), immunochemistry, and transcriptomic analyses demonstrated that the G-layer induced in flax xylem fibers was similar to the constitutively formed tertiary cell wall of bast (phloem) fibers but different from the secondary cell wall. The tertiary cell walls, independent of tissue of origin and inducibility, were twice as stiff as the secondary cell walls. In the gravitropic response, the tertiary cell wall deposition rate in xylem was higher than that of the secondary cell wall. Rhamnogalacturonan I (RG-I) with galactan side chains was a prominent component in cellulose-rich layers of both phloem and xylem flax fibers. Transcriptomic events underlying G-layer deposition in phloem and xylem fibers had much in common. At the induction of tertiary cell wall deposition, several genes for rhamnosyltransferases of the GT106 family were activated in xylem samples. The same genes were expressed in the isolated phloem fibers depositing the tertiary cell wall. The comparison of transcriptomes in fibers with both inducible and constitutive tertiary cell wall deposition and xylem tissues that formed the secondary cell walls is an effective system that revealed important molecular players involved in the formation of cellulose-enriched cell walls.

Auxin and gibberellin signaling cross-talk promotes hypocotyl xylem expansion and cambium homeostasis

During secondary growth, the thickening of plant organs, wood (xylem) and bast (phloem) is continuously produced by the vascular cambium. In Arabidopsis hypocotyl and root, we can distinguish two phases of secondary growth based on cell morphology and production rate. The first phase, in which xylem and phloem are equally produced, precedes the xylem expansion phase in which xylem formation is enhanced and xylem fibers differentiate. It is known that gibberellins (GA) trigger this developmental transition via degradation of DELLA proteins and that the cambium master regulator BREVIPEDICELLUS/KNAT1 (BP/KNAT1) and receptor like kinases ERECTA and ERL1 regulate this process downstream of GA. However, our understanding of the regulatory network underlying GA-mediated secondary growth is still limited. Here, we demonstrate that DELLA-mediated xylem expansion in Arabidopsis hypocotyl is mainly achieved through DELLA family members RGA and GAI, which promote cambium senescence. We further show that AUXIN RESPONSE FACTOR 6 (ARF6) and ARF8, which physically interact with DELLAs, specifically repress phloem proliferation and induce cambium senescence during the xylem expansion phase. Moreover, the inactivation of BP in arf6 arf8 background revealed an essential role for ARF6 and ARF8 in cambium establishment and maintenance. Overall, our results shed light on a pivotal hormone cross-talk between GA and auxin in the context of plant secondary growth.

AUXIN RESPONSE FACTOR 6 (ARF6) and ARF8 promote Gibberellin-mediated hypocotyl xylem expansion and cambium homeostasis

During secondary growth, the thickening of plant organs, wood (xylem) and bast (phloem) are continuously produced by the vascular cambium. In Arabidopsis hypocotyl and root, we can distinguish two phases of secondary growth based on cell morphology and production rate. The first phase, in which xylem and phloem are equally produced, precedes the xylem expansion phase in which xylem formation is enhanced and xylem fibers differentiate. It is known that Gibberellins (GA) trigger this developmental transition via the degradation of DELLA proteins and that the cambium master regulator BREVIPEDICELLUS/KNAT1 (BP/KNAT1) and the receptor like kinases ERECTA and ERL1 regulate this process downstream of GA. However, our understandings on the regulatory network underlying GA-mediated secondary growth, are still limited.Here, we demonstrate that DELLA-mediated xylem expansion is mainly achieved through RGA and GAI and that RGA and GAI promote cambium senescence. We further show that AUXIN RESPONSE FACTOR (ARF6) and ARF8, which physically interact with DELLAs, specifically repress phloem proliferation and induce cambium senescence during the xylem expansion phase. Moreover, the inactivation of BP in arf6 arf8 background revealed an essential role for ARF6 and ARF8 in cambium establishment and maintenance. Overall, our results shed light on a pivotal hormone cross-talk between GA and auxin in the context of plant secondary growth.

The in vivo impact of MsLAC1, a Miscanthus laccase isoform, on lignification and lignin composition contrasts with its in vitro substrate preference

Background Understanding lignin biosynthesis and composition is of central importance for sustainable bioenergy and biomaterials production. Species of the genus Miscanthus have emerged as promising bioenergy crop due to their rapid growth and modest nutrient requirements. However, lignin polymerization in Miscanthus is poorly understood. It was previously shown that plant laccases are phenol oxidases that have multiple functions in plant, one of which is the polymerization of monolignols. Herein, we link a newly discovered Miscanthus laccase, MsLAC1, to cell wall lignification. Characterization of recombinant MsLAC1 and Arabidopsis transgenic plants expressing MsLAC1 were carried out to understand the function of MsLAC1 both in vitro and in vivo. Results Using a comprehensive suite of molecular, biochemical and histochemical analyses, we show that MsLAC1 localizes to cell walls and identify Miscanthus transcription factors capable of regulating MsLAC1 expression. In addition, MsLAC1 complements the Arabidopsis lac4–2 lac17 mutant and recombinant MsLAC1 is able to oxidize monolignol in vitro. Transgenic Arabidopsis plants over-expressing MsLAC1 show higher G-lignin content, although recombinant MsLAC1 seemed to prefer sinapyl alcohol as substrate. Conclusions In summary, our results suggest that MsLAC1 is regulated by secondary cell wall MYB transcription factors and is involved in lignification of xylem fibers. This report identifies MsLAC1 as a promising breeding target in Miscanthus for biofuel and biomaterial applications.

The R2R3 MYB transcription factor MYB189 negatively regulates secondary cell wall biosynthesis in Populus

Secondary cell wall (SCW) biosynthesis during wood formation in trees is controlled by a multilevel regulatory network that coordinates the expression of substantial genes. However, few transcription factors involved in the negative regulation of secondary wall biosynthesis have been characterized in tree species. In this study, we isolated an R2R3 MYB transcription factor MYB189 from Populus trichocarpa, which is expressed predominantly in secondary vascular tissues, especially in the xylem. A novel repression motif was identified in the C-terminal region of MYB189, which indicates this factor was a transcriptional repressor. Overexpression (OE) of MYB189 in Arabidopsis and poplar resulted in a significant reduction in the contents of lignin, cellulose and hemicelluloses. Vascular development in stems of MYB189 OE lines was markedly inhibited, leading to a dramatic decrease in SCW thickness of xylem fibers. Gene expression analyses showed that most of the structural genes involved in the biosynthesis of lignin, cellulose and xylans were significantly downregulated in MYB189-overexpressing poplars compared with the wild-type control. Chromatin immunoprecipitation-quantitative real-time polymerase chain reaction and transient expression assays revealed that MYB189 could directly bind to the promoters of secondary wall biosynthetic genes to repress their expression. Together, these data suggest that MYB189 acts as a repressor to regulate SCW biosynthesis in poplar.

Development of successive cambia and structure of the secondary xylem in some members of the family Amaranthaceae

Young stems of Aerva javanica (Burm.f.) Juss. ex Schult., A. lanata (L.) Juss. ex Schult, A. monsonia Mart., A. sanguinolenta (L.) Blume, Alternanthera bettzickiana (Regel) G. Nicholson, A. philoxeroides (Mart.) Griseb., Gomphrena celosioides Mart., G. globosa L. and Telanthera ficoidea (L.) Moq., showed the renewal of small sectors of cambium by replacing with new segments. Therefore, the secondary phloem formed by earlier cambial segments form isolated islands of phloem enclosed within conjunctive tissues became embedded in the secondary xylem. As the stem grows older, complete ring of cambium is renewed; sometimes an anastomosing network of successive cambia may be seen due to the renewal of larger segments of the cambium. Renewal of the cambium takes place by repeated periclinal division in the parenchyma cells positioned outside to the phloem formed by the previous cambium. Functionally the cambium is bidirectional and exclusively composed of fusiform cambial cells. Differentiation of conducting elements of the secondary xylem and phloem remains restricted to the certain cambial cells while rest of the segments exclusively produce conjunctive cells. Accumulation of starch along with the presence of nuclei in the xylem fibers even after deposition of the secondary wall is consistent in all the species and it seems to be associated with the absence of rays in the secondary xylem and phloem of nine species from four genera. The significance of successive cambia, rayless xylem and nucleated xylem fibers were correlated with plant habit.